Method for forming a nanomedicinal composition

ABSTRACT

A nanomedicinal composition comprising a nanocarrier and an antioxidant. The nanocarrier contains a metal organic framework and a porous silicate and/or aluminosilicate matrix. The antioxidant is disposed in the pores and/or on the surface of the nanocarrier by a solution phase impregnation process. The nanomedicinal composition is used in a method of treating Blastocystis infection.

BACKGROUND OF THE INVENTION Field of the Invention

The present disclosure relates to a nanomedicinal composition comprisinga nanocarrier and an antioxidant, a method of its preparation, and amethod of treating an infection by parasite in the genus Blastocystis.

Discussion of the Background

The “background” description provided herein is for the purpose ofgenerally presenting the context of the disclosure. Work of thepresently named inventors, to the extent it is described in thisbackground section, as well as aspects of the description which may nototherwise qualify as prior art at the time of filing, are neitherexpressly nor impliedly admitted as prior art against the presentinvention.

Parasitic diseases are a major public health threat in Saudi Arabia andworldwide. These diseases are responsible for considerable morbidity andmortality. Blastocystis is a unicellular parasitic protozoon. Itinhabits the large intestine of humans and animals, and is detectedglobally, with up to 100% prevalence [El Safadi, D., et. al., BMC InfectDis., 2016, 26, 451]. Blastocystosis, human infection by Blastocystis,has various outcomes: it can be asymptomatic in infected patients, canproduce diarrhea and other gastrointestinal symptoms, or have anopportunistic character and cause infection in immunocompromisedpatients. Blastocystis has been linked to symptoms of inflammatory boweldiseases, irritable bowel syndrome and acute urticaria[Katsarou-Katsari, A., et. al., Acta Derm Venereo. 2008, 88, 80-81; &Roberts, T., et. al., Gut Pathogens, 2014, 6, 1, 17]. Recently,Blastocystis was linked to gut dysbiosis and associated gut disorders aswell as the induction of growth of colon and rectum cancer by apoptosisof cancer colon cells [Tan, T. C., et. al., Parasitol Res., 2009, 105,1283-1286; Chandramathi, S., et. al., Parasitol Res., 2020, 106, 4,941-945; & Stensvold, C. R., et. al., Trends Parasitol., 2020, 3,315-316].

Drugs currently used to treat Blastocystis are not fully effective.Among the many drugs used to treat Blastocystis infection, metronidazoleis the most effective therapy, however, there remains controversy aboutresponses of different categories of diarrheic patients in addition toside effects and drug resistance [Rajamanikam, A., et. al., PLoS One,2019, 14, 2, e0212542]. Some antioxidant compounds have exhibitedactivity against Blastocistis, but the bioavailability of most effectiveantioxidant compounds is too low to be clinically useful. Consequently,there is a vital need to develop effective alternative therapies totreat and control this disease.

Accordingly, it is an objective of the present disclosure to provide ananocarrier for certain antioxidant compounds with the aim of providinga method of treating Blastocystis infection.

SUMMARY OF THE INVENTION

The present disclosure relates to a nanomedicinal composition,comprising a nanocarrier comprising a metal organic framework which is azeolitic imidazolate framework, and a porous silicate and/oraluminosilicate matrix, and an antioxidant disposed in the pores and/oron a surface of the nanocarrier.

In some embodiments, the porous silicate and/or aluminosilicate matrixis at least one selected from the group consisting of MCM-41 and KIT-6.

In some embodiments, the nanocarrier has a surface area of 225 to 750m²/g, a pore volume of 0.25 to 0.85 cm³/g, and a mean pore size of 2 to10 nm.

In some embodiments, the porous silicate and/or aluminosilicate matrixis MCM-41 and the nanocarrier has a surface area of 450 to 750 m²/g, apore volume of 0.25 to 0.65 cm³/g, and a mean pore size of 2 to 4 nm.

In some embodiments, the porous silicate and/or aluminosilicate matrixis KIT-6 and the nanocarrier has a surface area of 225 to 450 m²/g, apore volume of 0.45 to 0.85 cm³/g, and a mean pore size of 5 to 10 nm.

In some embodiments, the porous silicate and/or aluminosilicate matrixis present in an amount of 1 to 20 wt % based on a total weight of thenanocarrier.

In some embodiments, the metal organic framework comprises an imidazoleof formula I:

and is substantially free of a benzimidazole of formula II:

In some embodiments, the zeolitic imidazolate framework is ZIF-8.

In some embodiments, the antioxidant is at least one selected from thegroup consisting of quercetin, rutin, coenzyme Q10, gallic acid,resveratrol, and curcumin.

In some embodiments, the antioxidant is curcumin.

In some embodiments, the antioxidant is resveratrol.

In some embodiments, the antioxidant is present in the nanomedicinalcomposition in an amount of 5 to 50 wt %, based on a total weight ofnanomedicinal composition.

In some embodiments, the nanomedicinal composition releases greater than20% of a total weight of curcumin within 24 to 72 hours of contact witha suitable biological medium.

In some embodiments, the nanomedicinal composition releases greater than7.5% of a total weight of resveratrol within 24 to 72 hours of contactwith a suitable biological medium.

In some embodiments, the nanomedicinal composition reduces the viabilityof Blastocystis organisms by at least 75% when the nanomedicinalcomposition is present for 24 hours in an amount of 100 to 1000 μg/mL.

The present disclosure also relates to a method of forming thenanomedicinal composition comprising mixing the metal organic frameworkand the porous silicate and/or aluminosilicate matrix to form thenanocarrier, combining the nanocarrier and the antioxidant in animpregnation solution thereby forming the nanomedicinal composition, andisolating the nanomedicinal composition.

In some embodiments, the mixing comprises ultrasonication.

In some embodiments, the impregnation solution comprises an alcoholhaving 1 to 5 carbon atoms and the antioxidant is present in an amountof 0.5 to 3 mg/mL of impregnation solution.

The present disclosure also relates to a method for treating aninfection by a parasite in the genus Blastocystis in a subject,comprising administering to a subject in need of therapy apharmaceutical composition comprising the nanomedicinal composition.

The present disclosure also relates to a pharmaceutical composition,comprising the nanomedicinal composition of claim 1 and apharmaceutically acceptable excipient.

BRIEF DESCRIPTION OF THE DRAWINGS

A more complete appreciation of this disclosure and many of theattendant advantages thereof will be readily obtained as the samebecomes better understood by reference to the following detaileddescription when considered in connection with the accompanyingdrawings, wherein:

FIG. 1 shows a schematic overview of the present invention.

FIG. 2 shows X-ray diffraction pattern of ZIF-8, Resveratrol, Curcumin,MCM-41/ZIF-8/Curcumin, KIT-6/ZIF-8/Curcumin, MCM-41/ZIF-8/Resveratrol,and KIT-6/ZIF-8/Resveratrol nanocomposites.

FIG. 3A shows nitrogen adsorption isotherms of MCM-41, MCM-41/ZIF-8,KIT-6, and KIT-6/ZIF-8.

FIG. 3B shows a pore size distribution plot of MCM-41, MCM-41/ZIF-8,KIT-6, and KIT-6/ZIF-8.

FIGS. 4A-4B show transmission electron microscope (TEM) images ofKIT-6/ZIF-8 at magnification of 200 nm (FIG. 4A) and 20 nm (FIG. 4B).

FIG. 5A shows the FTIR spectra of Resveratrol, ZIF-8, MCM-41,MCM-41/ZIF-8/Resveratrol, and KIT-6/ZIF-8/Resveratrol nanocomposites.

FIG. 5B shows the FTIR spectra of Curcumin, MCM-41/ZIF-8/Curcumin, andKIT-6/ZIF-8/Curcumin nanocomposites.

FIG. 6 shows the diffuse reflectance spectra of KIT-6/ZIF-8,Resveratrol, Curcumin, MCM-41/ZIF-8/Curcumin, KIT-6/ZIF-8/Curcumin,MCM-41/ZIF-8/Resveratrol, and KIT-6/ZIF-8/Resveratrol nanocomposites.

FIG. 7 shows the release profile of antioxidants loaded nanocompositesat 37° C. for 96 h Resveratrol (pH=5.6), MCM-41/ZIF-8//Resveratrol(pH=5.6), MCM-41/ZIF-8//Resveratrol (pH=7.4), KIT-6/ZIF-8/Resveratrol(pH=5.6), KIT-6/ZIF-8/Resveratrol (pH=7.4), KIT-6/Curcumin (pH=5.6),KIT-6/ZIF-8/Curcumin (pH=5.6), and MCM-41/ZIF-8/Curcumin (pH=5.6).

FIGS. 8A-8C show the reduction in the percentage of Blastocystis cystsviability after exposure to increasing concentrations ofresveratrol/ZIF-8/Kit-6 nanocomposite (FIG. 8A), curcumin/ZIF-8/MCM-41nanocomposite (FIG. 8B), and ZIF-8/KIT-6 nanocomposite (FIG. 8C).

DETAILED DESCRIPTION OF THE INVENTION

In the following description, it is understood that other embodimentsmay be utilized and structural and operational changes may be madewithout departure from the scope of the present embodiments disclosedherein.

Definitions

Embodiments of the present disclosure will now be described more fullyhereinafter with reference to the accompanying drawings, in which some,but not all embodiments of the disclosure are shown.

As used herein, the words “a” and “an” and the like carry the meaning of“one or more”. Within the description of this disclosure, where anumerical limit or range is stated, the endpoints are included unlessstated otherwise. Also, all values and subranges within a numericallimit or range are specifically included as if explicitly written out.

As used herein, the terms “optional” or “optionally” means that thesubsequently described event(s) can or cannot occur or the subsequentlydescribed component(s) may or may not be present (e.g., 0 wt. %).

The phrase “substantially free”, unless otherwise specified, describesan amount of a particular component (e.g., a benzimidazole of Formula(II)), that when present, is present in an amount of less than about 1wt. %, preferably less than about 0.5 wt. %, more preferably less thanabout 0.1 wt. %, even more preferably less than about 0.05 wt. %,relative to a total weight of the composition being discussed, and alsoincludes situations where the composition is completely free of theparticular component (i.e., 0% wt.).

As used herein, the term “alkyl” unless otherwise specified refers toboth branched and straight chain saturated aliphatic primary, secondary,and/or tertiary hydrocarbon fragments of typically C₁ to C₂₀.Non-limiting examples of such hydrocarbon fragments include methyl,trifluoromethyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl,t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl,cyclohexyl, cyclohexylmethyl, 3-methylpentyl, 2,2-dimethylbutyl,2,3-dimethylbutyl, 2-ethylhexyl, heptyl, octyl, nonyl,3,7-dimethyloctyl, decyl, undecyl, dodecyl, tridecyl, 2-propylheptyl,tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, andeicosyl.

The term “cycloalkyl” refers to cyclized alkyl groups. Exemplarycycloalkyl groups include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, and adamantyl. Branchedcycloalkyl groups such as exemplary 1-methylcyclopropyl and2-methylcyclopropyl groups are included in the definition of cycloalkylas used in the present disclosure.

The term “alkoxy” refers to a straight or branched chain alkoxyincluding, but not limited to, methoxy, ethoxy, propoxy, isopropoxy,butoxy, isobutoxy, secondary butoxy, tertiary butoxy, pentoxy,isopentoxy, hexyloxy, heptyloxy, octyloxy, nonyloxy, and decyloxy.

The term “halogen”, as used herein, means fluoro, chloro, bromo andiodo.

As used herein, the term “substituted” refers to at least one hydrogenatom is replaced with a non-hydrogen group, provided that normalvalencies are maintained and that the substitution results in a stablecompound. When a R group (denoted as R₁, R₂, and so forth) is noted as“optionally substituted”, the substituents are selected from theexemplary group including, but not limited to, halogen (e.g., chlorine,bromine, fluorine or iodine), alkoxy (i.e., straight chain alkoxy having1 to 3 carbon atoms, and includes, for example, methoxy, ethoxy, andpropoxy), hydroxy, amino, alkylamino, thiol, alkylthio, sulfonamido(e.g., —SO₂NH₂), substituted sulfonamide (e.g., —SO₂ NHalkyl or caseswhere there are two alkyl substituents on one nitrogen), nitro, cyano,carboxy, carbamyl (e.g., —CONH₂), substituted carbamyl (e.g., —CONHalkylor cases where there are two alkyl substituents on one nitrogen), andmixtures thereof. The substituents may be either unprotected, orprotected as necessary, as known to those skilled in the art, forexample, as taught in Greene et al., “Protective Groups in OrganicSynthesis”, John Wiley and Sons, Second Edition, 1991, herebyincorporated by reference in its entirety).

According to a first aspect, the present disclosure relates to ananomedicinal composition, comprising a nanocarrier comprising a metalorganic framework, and a porous silicate and/or aluminosilicate matrix,and an antioxidant disposed in the pores and/or on a surface of thenanocarrier.

The International Union of Pure and Applied Chemistry (IUPAC) statesthat a metal organic framework (MOF) is a coordination network withorganic ligands containing potential voids. A coordination network is acoordination compound extending, through repeating coordinationentities, in one dimension, but with cross-links between two or moreindividual chains, loops, or spiro-links, or a coordination compoundextending through repeating coordination entities in two or threedimensions; and finally a coordination polymer is a coordinationcompound with repeating coordination entities extending in one, two, orthree dimensions. A coordination entity is an ion or neutral moleculethat is composed of a central atom, usually that of a metal, to which isattached a surrounding array of atoms or groups of atoms, each of whichis called ligands. More succinctly, a metal organic framework ischaracterized by metal ions or clusters coordinated to organic ligandsto form one-, two-, or three-dimensional structures. Typically, a MOFexhibits a regular void or pore structure. The nature of the void orpore structure, including properties or structural factors such as thegeometry about the metal ions or clusters, the arrangement of thelinkages between metal ions or clusters, and the number, identity, andspatial arrangement of voids or pores. These properties may be describedas the structure of the repeat units and the nature of the arrangementof the repeat units. The specific structure of the MOF, which mayinclude the void or pore structure is typically referred to as the MOFtopology.

The metal-organic framework comprises a metal ion which is an ion of atleast one metal selected from the group consisting of a transition metal(e.g. Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Y, Zr, Nb, Mo, Tc, Ru, Rh, Pd,Ag, Hf, Ta, W, Re, Os, Ir, Pt, Au, Rf, Db, Sg, Bh, Hs, Mt, Ds, Rg, andCn), a post-transition metal (e.g. Al, In, Ga, Sn, Bi, Pb, Tl, Zn, Cd,and Hg), and an alkaline earth metal (e.g. Be, Mg, Ca, Sr, Ba, and Ra).Further, these metal ions may be of any oxidation state M⁺¹, M⁺², M⁺³,etc. In one or more embodiments, the metal ion is an ion of at least onemetal selected from the group consisting of Zn, Cu, Fe, Ni, Co, Mn, Cr,Cd, Mg, Ca, and Zr. In a preferred embodiment, the at least one metal isZn.

In the formation of a metal organic framework, the organic ligands mustmeet certain requirements to form coordination bonds, primarily beingmulti-dentate, having at least two donor atoms (i.e. N—, and/or O—) andbeing neutral or anionic. The structure of the metal organic frameworkis also affected by the shape, length, and functional groups present inthe organic linker. In certain embodiments, the metal organic frameworkof the present disclosure comprises anionic ligands as organic ligands.In one or more embodiments, the organic ligands may have at least twonitrogen donor atoms. For example, the organic ligands may beimidazolate-based, imidazole-derived or ligands similar to an imidazoleincluding, but not limited to, optionally substituted imidazoles,optionally substituted benzimidazoles, optionally substitutedimidazolines, optionally substituted pyrazoles, optionally substitutedthiazoles, and optionally substituted triazoles. In a preferredembodiment, the metal organic framework of the present disclosure in anyof its embodiments comprises 2-methylimidazole and 5-methylbenzimidazoleas the organic ligands. 2-Methylimidazole and 5-methylbenzimidazoleorganic ligands have free nitrogen atoms that may each form acoordinative bond to the metal ions (e.g. Zn(II)) to produce acoordination network.

In one or more embodiments, the ligand comprises an imidazole of formula(I) and/or a benzimidazole of formula (II):

wherein R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are each independentlyselected from the group consisting of a hydrogen, an optionallysubstituted alkyl, an optionally substituted cycloalkyl, an optionallysubstituted alkoxy, a hydroxyl, a halogen, a nitro, and a cyano.Preferably, R₁, R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are each independently ahydrogen, an optionally substituted C₁-C₃ alkyl group, or an optionallysubstituted C₃-C₆ cycloalkyl group. More preferably, R₁, R₂, R₃, R₄, R₅,R₆, R₇, and R₈ are each independently a hydrogen or a methyl.

Exemplary imidazole-based ligands that may be applicable to the currentdisclosure include, but are not limited to, imidazole,2-methylimidazole, 4-methylimidazole, 2-ethylimidazole,2-isopropylimidazole, 4-tert-butyl-1H-imidazole,2-ethyl-4-methylimidazole, 2-bromo-1H-imidazole, 4-bromo-1H-imidazole,2-chloro-1H-imidazole, 2-iodoimidazole, 2-nitroimidazole,4-nitroimidazole, (1H-imidazol-2-yl)methanol,4-(hydroxymethyl)imidazole, 2-aminoimidazole,4-(trifluoromethyl)-1H-imidazole, 4-cyanoimidazole,3H-imidazole-4-carboxylic acid, 4-imidazolecarboxylic acid,imidazole-2-carboxylic acid, 2-hydroxy-1H-imidazole-4-carboxylic acid,4,5-imidazoledicarboxylic acid, 5-iodo-2-methyl-1H-imidazole,2-methyl-4-nitroimidazole, 2-(aminomethyl)imidazole, 4,5-dicyanoimidazole, 4-imidazoleacetic acid, 4-methyl-5-imidazolemethanol,1-(4-methyl-1H-imidazol-5-yl)methanamine, 4-imidazoleacrylic acid,5-bromo-2-propyl-1H-imidazole, ethyl-(1H-imidazol-2-ylmethyl)-amine, and2-butyl-5-hydroxymethylimidazole. In preferred embodiments, theimidazole of formula (I) is 2-methylimidazole.

Exemplary benzimidazole-based ligands that may be applicable to thecurrent disclosure include, but are not limited to, benzimidazole,5-methylbenzimidazole, 2-methylbenzimidazole, 5-chlorobenzimidazole,5-bromobenzimidazole, 5,6-dimethylbenzimidazole, 5-methoxybenzimidazole,2-chlorobenzimidazole, 2-bromo-1H-benzimidazole,6-bromo-1H-benzimidazole, 5-fluoro-1H-benzimidazole,5-chloro-2-methylbenzimidazole, methyl benzimidazole-2-acetate,1H-benzoimidazol-4-ol, 1H-benzimidazol-5-ylmethanol,2-benzimidazolemethanol, 4-chloro-6-(trifluoromethyl)benzimidazole,5-chloro-2-(trichloromethyl)benzimidazole, 5-cyanobenzimidazole,(2-benzimidazolyl)acetonitrile, (5-chloro-1H-benzimidazol-2-yl)methanol,2-(chloromethyl)benzimidazole, 5-iodo-2-methylbenzimidazole,(5-chloro-1H-benzimidazol-2-yl)methylamine,2-(aminomethyl)benzimidazole, 2-(6-chloro-1H-benzimidazol-2-yl)ethanol,2-(1H-benzoimidazol-2-yl)-acetamide,(6-methoxy-1H-benzimidazol-2-yl)methanol, 5,6-dimethoxybenzimidazole,2-(1H-benzoimidazol-2-yl)-ethylamine,1-(5-methyl-1H-benzimidazol-2-yl)methanamine,1-(5-methyl-1H-benzimidazol-2-yl)ethanamine, 2-benzimidazolepropionicacid, 2-(5-methyl-1H-benzimidazol-2-y1)ethanamine,2-(3-hydroxy-N-propyl)-5-(trifluoromethyl)-benzimidazole, andN-methyl-1-(5-methyl-1H-benzimidazol-2-yl)methanamine. In someembodiments, the benzimidazole of formula (II) is 5-methylbenzimidazole.

In one or more embodiments, the metal organic framework comprises aimidazole of formula (I). In one or more embodiments, the metal organicframework is substantially free of a benzimidazole of formula (II).

Metal organic frameworks comprising such imidazole or benzimidazoleligands are typically referred to as zeolitic imidazolate frameworks. Insome embodiments, the metal organic framework is a zeolitic imidazolateframework. In one or more embodiments, the metal-organic frameworkcomprises ZIF-8. In preferred embodiments, the metal-organic frameworkis ZIF-8. Other metal-organic frameworks that may be used in thecurrently disclosed membrane include, but are not limited to,isoreticular metal organic framework-3 (IRMOF-3), MOF-69A, MOF-69B,MOF-69C, MOF-70, MOF-71, MOF-73, MOF-74, MOF-75, MOF-76, MOF-77, MOF-78,MOF-79, MOF-80, DMOF-1-NH2, UMCM-1-NH2, MOF-69-80, ZIF-1, ZIF-2, ZIF-3,ZIF-4, ZIF-5, ZIF-6, ZIF-7, ZIF-9, ZIF-10, ZIF-11, ZIF-12, ZIF-14,ZIF-20, ZIF-21, ZIF-22, ZIF-23, ZIF-25, ZIF-60, ZIF-61, ZIF-62, ZIF-63,ZIF-64, ZIF-65, ZIF-66, ZIF-67, ZIF-68, ZIF-69, ZIF-70, ZIF-71, ZIF-72,ZIF-73, ZIF-74, ZIF-75, ZIF-76, ZIF-77, ZIF-78, ZIF-79, ZIF-80, ZIF-81,ZIF-82, ZIF-90, ZIF-91, ZIF-92, ZIF-93, ZIF-94, ZIF-96, ZIF-97, ZIF-100,ZIF-108, ZIF-303, ZIF-360, ZIF-365, ZIF-376, ZIF-386, ZIF-408, ZIF-410,ZIF-412, ZIF-413, ZIF-414, ZIF-486, ZIF-516, ZIF-586, ZIF-615, andZIF-725.

In some embodiments, the metal organic framework is present in the formof particles. In general, the metal organic framework particles can beany shape known to one of ordinary skill in the art. Examples ofsuitable shapes the metal organic framework particles may take includespheres, spheroids, lentoids, ovoids, solid polyhedra such astetrahedra, cubes, octahedra, icosahedra, dodecahedra, rectangularprisms, triangular prisms (also known as nanotriangles), nanoplatelets,nanodisks, blocks, flakes, discs, granules, angular chunks, and mixturesthereof. Nanorods or nanowires are not a shape that the metal organicframework particles are envisioned as having in any embodiments.

In some embodiments, the metal organic framework particles have uniformshape. Alternatively, the shape may be non-uniform. As used herein, theterm “uniform shape” refers to an average consistent shape that differsby no more than 10%, by no more than 5%, by no more than 4%, by no morethan 3%, by no more than 2%, by no more than 1% of the distribution ofmetal organic framework particles having a different shape. As usedherein, the term “non-uniform shape” refers to an average consistentshape that differs by more than 10% of the distribution of metal organicframework particles having a different shape. In one embodiment, theshape is uniform and at least 90% of the metal organic frameworkparticles are spherical or substantially circular, and less than 10% arepolygonal. In another embodiment, the shape is non-uniform and less than90% of the metal organic framework particles are spherical orsubstantially circular, and greater than 10% are polygonal.

In some embodiment, the metal organic framework is in the form ofparticles having a mean particle size of 50 to 10,000 nm, preferably 75to 9,000 nm, preferably 100 to 8,000 nm, preferably 125 to 7,500 nmpreferably 150 to 7,000 nm. In embodiments where the metal organicframework particles are spherical, the particle size may refer to aparticle diameter. In embodiments where the metal organic frameworkparticles are polyhedral, the particle size may refer to the diameter ofa circumsphere. In some embodiments, the particle size refers to a meandistance from a particle surface to particle centroid or center of mass.In alternative embodiments, the particle size refers to a maximumdistance from a particle surface to a particle centroid or center ofmass. In some embodiments where the metal organic framework particleshave an anisotropic shape such as nanorods or nanotubes, the particlesize may refer to a length of the nanorod or nanotube, a width of thenanorod or nanotube, or an average of the length and width of thenanorod or nanotube. In some embodiments, the particle size refers tothe diameter of a sphere having an equivalent volume as the particle.

In general, any suitable silicate and/or aluminosilicate matrix known toone of ordinary skill in the art may be used in the nanomedicinalcomposition. Examples of such suitable porous silica, silicate, oraluminosilicate materials include, but are not limited to, MCM-41,MCM-48, Q-10 silica, hydrophobic silica, mesobeta, mesoZSM-5, SBA-15,KIT-5, KIT-6, mesosilicalite, hierarchical porous silicalite, SBA-16,mesoporous silica spheres, and halloysite. The term “silicate matrix”should be understood to include silica itself. Methods of obtaining thevarious types porous silica, silicate, or aluminosilicate material arewell-known in the art [see for example Gobin, Oliver Christian “SBA-16Materials: Synthesis, Diffusion, and Sorption Properties” Dissertation,Laval University, Ste-Foy, Quebec, Canada, January 2006, in particularsection 2.2; and U.S. patent application Ser. No. 15/478,794—bothincorporated herein by reference in their entireties]. Aluminosilicatematerials may be characterized by a ratio of Si to Al present in thematerial. In general, the aluminosilicate material may have any suitableSi:Al molar ratio. Examples of such suitable Si:Al molar ratios are1000:1 to 1:250, preferably 500:1 to 1:200, preferably 250:1 to 1:100,preferably 150:1 to 1:75, preferably 100:1 to 1:50, preferably 50:1 to1:25, preferably 25:1 to 1:10, preferably 10:1 to 1:5, preferably 5:1 to1:2.5, preferably 2.5:1 to 1:1.5, preferably 1.5:1 to 1:1. In general,the elemental composition of the silicate and/or aluminosilicatematerial, including the Si:Al molar ratio, may be determined by anysuitable technique known to one of ordinary skill in the art. Examplesof suitable such techniques include mass spectrometry techniques such asinductively-coupled plasma mass spectrometry (ICP-MS), atomic emissionspectroscopy techniques such as inductively-coupled plasma atomicemission spectroscopy (ICP-AES) (also referred to as ICP opticalemission spectroscopy, ICP-OES), atomic absorption spectroscopytechniques such as inductively-coupled plasma atomic absorptionspectroscopy (ICP-AAS), and X-ray spectroscopy techniques such as X-rayphotoelectron spectroscopy.

Silicates and aluminosilicates are materials which comprise SiO₄tetrahedra (and AlO₄ ⁻ tetrahedra, AlO₆ octahedra, and/or Al(OH)₆octahedra in the case of aluminosilicates) joined together in a widevariety of structural motifs. The tetrahedra (and if applicableoctahedra) in the silicate and/or aluminosilicate material of thepresent invention may in general adopt any structural motif present inother silicate materials, such as isolated tetradhedra as inneosilicates (single tetrahedra, also called orthosilicates) andsorosilicates (double tetrahedra), chains of tetrahedra such asinosilicates (both single chain as in pyroxene group silicates anddouble chain as in amphibole group silicates), rings of tetrahedra as incyclosilicates, sheets of tetrahedra as in phyllosilicates, andthree-dimensional frameworks as in tectosilicates. In somealuminosilicates, the material comprises a substructure comprisingsilicon-containing and/or aluminum-containing tetrahedral and asubstructure comprising aluminum-containing octahedral. An example ofsuch an arrangement is the mineral kaolin, which comprises sheets ofalternating tetrahedra-containing layers and octahedra-containinglayers. The arrangement of isolated tetrahedra, chains of tetrahedra,sheets of tetrahedra, or three-dimensional frameworks may give rise tochannels, pores, cages, or other spaces within the silicate and/oraluminosilicate which is capable of hosting material which is not thesilicate and/or aluminosilicate itself. Examples of materials,particularly those relevant to the current disclosure, include water,organic molecules, and inorganic nanoparticles. While the largerstructures formed of tetrahedra (i.e. chains, rings, sheets, andthree-dimensional frameworks) may themselves be ordered, the arrangementof these larger structures may be disordered. Such disorder may giverise to a material which is amorphous by techniques for determiningcrystallinity or crystal structure such as powder X-ray diffraction(PXRD). Alternatively, the larger structures may be ordered, giving riseto a crystalline material.

MCM-41 (Mobil Composition of Matter No. 41) is a mesoporous silicamaterial with a hierarchical structure from a family of silicate andaluminosilicate solids that were developed by researchers at Mobil OilCorporation and that can be used as catalysts or catalyst supports.MCM-41 and MCM-48 both comprise an amorphous silica wall and possesslong range ordered framework with uniform mesopores. These materialsalso possess large surface area, which can be up to more than 1,000m²g⁻¹. The pore diameter of these materials can be controlled to fallwithin a mesoporous range between 1.5 and 20 nm by adjusting thesynthesis conditions and/or by employing surfactants with differentchain lengths in their preparation. In embodiments where the poroussilicate matrix is MCM-41, the nanocarrier may be referred to as a“MCM-41 nanocarrier”.

KIT-6 is a mesoporous silica material. KIT-6 has a bicontinuous cubicmesostructure with Ia3d symmetry. KIT-6 is characterized by aninterpenetrating cylindrical pore system. Such pores typically have apore size from about 3.5 to about 18.5 nm and can be controlled byvarious parameters during the synthesis such as synthesis temperature.

In some embodiments, the porous silicate and/or aluminosilicate matrixis surface modified prior to use in the nanocarrier. Such surfacemodifications may change the surface properties of the porous silicateand/or aluminosilicate matrix, for example by increasing or decreasingthe number or concentration of functional groups found on an unmodifiedporous silicate and/or aluminosilicate matrix or by introducing newfunctional groups to the porous silicate and/or aluminosilicate matrix.Examples of such new functional groups include, but are not limited tocarboxylic acid or carboxylate groups, amine or ammonium groups, sulfogroups, and phosphate groups. Such functional groups may be charged oruncharged. In some embodiments, the surface modification changes thesurface charge of the interior surface, the exterior surface, the poresurface, or any combination thereof of the modified porous silicateand/or aluminosilicate matrix compared to unmodified porous silicateand/or aluminosilicate matrix. Preferably, the surface modification doesnot change the surface charge of the interior surface, exterior surface,pore surface, or any combination thereof of the modified porous silicateand/or aluminosilicate matrix compared to unmodified porous silicateand/or aluminosilicate matrix. Such surface modification may beperformed using any suitable method or with any suitable surfacemodifying agent or agents known to one of ordinary skill in the art. Oneexample of such a method is the use of silanes or organosilicatesbearing one or more functional groups to be added by the surfacemodification. Such surface modification may result in said functionalgroups being attached to the porous silicate and/or aluminosilicatematrix by covalent bonds. Alternatively, said functional groups may beattached to the porous silicate and/or aluminosilicate matrix by anon-covalent interaction, for example electrostatic interaction,physisorption, or hydrogen bonding. In some embodiments, the surfacemodifying agent(s) are substantially free of silianes. In someembodiments, the surface modifying agent(s) are substantially free oforganosilicates. In some embodiments, the surface modifying agent(s) aresubstantially free of amino acids. In some embodiments, the surfacemodifying agent(s) are substantially free of short peptides (i.e. 2-20residues). In some embodiments, the surface modifying agent(s) aresubstantially free of chromium salts (chrome alum, chromium acetate,etc.); calcium salts (calcium chloride, calcium hydroxide, etc.);aluminum salts (aluminum chloride, aluminumhydroxide, etc.); dialdehydes(glutaraldehyde, etc.); carbodiimides (EDC, WSC,N-hydroxy-5-norbomene-2,3-di-carboxylmide (HONB), N-hydroxysuccinic acidimide (HOSu), dicyclohexylcarbodiimide (DCC), etc.);N-hydrox-ysuccinimide; and/or phosphorus oxychloride. In someembodiments, the surface modifying agent(s) are substantially free ofproteins. Examples of such proteins include, but are not limited tocollagen, gelatin, albumin, ovalbumin, casein, transferrin, fibrin, andfibrinogen.

In some embodiments, the porous silicate and/or aluminosilicate matrixis present in the form of particles. In general, the porous silicateand/or aluminosilicate matrix particles can be any shape known to one ofordinary skill in the art. Examples of suitable shapes the poroussilicate and/or aluminosilicate matrix particles may take includespheres, spheroids, lentoids, ovoids, solid polyhedra such astetrahedra, cubes, octahedra, icosahedra, dodecahedra, rectangularprisms, triangular prisms (also known as nanotriangles), nanoplatelets,nanodisks, blocks, flakes, discs, granules, angular chunks, and mixturesthereof. Nanorods or nanowires are not a shape that the porous silicateand/or aluminosilicate matrix particles are envisioned as having in anyembodiments.

In some embodiments, the porous silicate and/or aluminosilicate matrixparticles have uniform shape. Alternatively, the shape may benon-uniform. As used herein, the term “uniform shape” refers to anaverage consistent shape that differs by no more than 10%, by no morethan 5%, by no more than 4%, by no more than 3%, by no more than 2%, byno more than 1% of the distribution of porous silicate and/oraluminosilicate matrix particles having a different shape. As usedherein, the term “non-uniform shape” refers to an average consistentshape that differs by more than 10% of the distribution of poroussilicate and/or aluminosilicate matrix particles having a differentshape. In one embodiment, the shape is uniform and at least 90% of theporous silicate and/or aluminosilicate matrix particles are spherical orsubstantially circular, and less than 10% are polygonal. In anotherembodiment, the shape is non-uniform and less than 90% of the poroussilicate and/or aluminosilicate matrix particles are spherical orsubstantially circular, and greater than 10% are polygonal.

In some embodiment, the porous silicate and/or aluminosilicate matrix isin the form of particles having a mean particle size of 50 to 10,000 nm,preferably 75 to 9,000 nm, preferably 100 to 8,000 nm, preferably 125 to7,500 nm preferably 150 to 7,000 nm. In embodiments where the poroussilicate and/or aluminosilicate matrix particles are spherical, theparticle size may refer to a particle diameter. In embodiments where theporous silicate and/or aluminosilicate matrix particles are polyhedral,the particle size may refer to the diameter of a circumsphere. In someembodiments, the particle size refers to a mean distance from a particlesurface to particle centroid or center of mass. In alternativeembodiments, the particle size refers to a maximum distance from aparticle surface to a particle centroid or center of mass. In someembodiments where the porous silicate and/or aluminosilicate matrixparticles have an anisotropic shape such as nanorods or nanotubes, theparticle size may refer to a length of the nanorod or nanotube, a widthof the nanorod or nanotube, or an average of the length and width of thenanorod or nanotube. In some embodiments, the particle size refers tothe diameter of a sphere having an equivalent volume as the particle.

In some embodiments, the porous silicate and/or aluminosilicate matrixis present in an amount of 1 to 20 wt %, preferably 2 to 18 wt %,preferably 3 to 17 wt %, preferably 4 to 16 wt %, preferably 5 to 15 wt%, preferably 6 to 14 wt %, preferably 7 to 13 wt %, preferably 8 to 12wt %, preferably 9 to 11 wt %, preferably 10 wt %, based on a totalweight of the nanocarrier.

In some embodiments, the nanocarrier has a surface area of 225 m²/g to750 m²/g, preferably 250 m²/g to 725 m²/g, preferably 275 m²/g to 700m²/g, preferably 300 m²/g to 675 m²/g. In some embodiments, the poroussilicate and/or aluminosilicate matrix is particles of MCM-41 and thenanocarrier has a surface area of 450 to 750 m²/g, preferably 460 to 740m²/g, preferably 470 to 730 m²/g, preferably 480 to 720 m²/g, preferably490 to 710 m²/g, preferably 500 to 700 m²/g, preferably 510 to 690 m²/g,preferably 520 to 680 m²/g, preferably 530 to 670 m²/g, preferably 540to 660 m²/g, preferably 550 to 650 m²/g, preferably 560 to 640 m²/g,preferably 570 to 630 m²/g, preferably 580 to 620 m²/g, preferably 590to 610 m²/g. In some embodiments, the porous silicate and/oraluminosilicate matrix is particles of KIT-6 and the nanocarrier has asurface area of 225 to 450 m²/g, preferably 235 to 440 m²/g, preferably245 to 430 m²/g, preferably 255 to 420 m²/g, preferably 265 to 410 m²/g,preferably 275 to 400 m²/g, preferably 285 to 390 m²/g, preferably 295to 380 m²/g, preferably 305 to 370 m²/g, preferably 315 to 360 m²/g,preferably 325 to 350 m²/g, preferably 335 to 340 m²/g.

In some embodiments, the nanocarrier has a mean pore size of 1 nm to 60nm, preferably 1.25 to 50 nm, preferably 1.5 nm to 40 nm, preferably1.75 nm to 30 nm, preferably 2 nm to 20 nm, preferably 2.25 to 10 nm,preferably 2.5 to 9 nm, preferably 2.75 to 8 nm, preferably 3.0 to 7.75nm. In some embodiments, the porous silicate and/or aluminosilicatematrix is particles of MCM-41 and the nanocarrier has a mean pore sizeof 2 to 4 nm, preferably 2.25 to 3.75 nm, preferably 2.5 to 3.5 nm,preferably 2.75 to 3.25 nm, preferably 2.9 to 3.1 nm, preferably 3 nm.In some embodiments, the porous silicate and/or aluminosilicate matrixis particles of KIT-6 and the nanocarrier has a mean pore size of 5 to10 nm, preferably 5.25 to 9.75 nm, preferably 5.5 to 9.5 nm, preferably6 to 9 nm, preferably 6.25 to 8.75 nm, preferably 6.5 to 8.5 nm,preferably 6.75 to 8.25 nm, preferably 7 to 8 nm, preferably 7.25 to7.75 nm, preferably 7.5 to 7.65 nm.

In some embodiments, the nanocarrier has a mean pore volume of 0.25 to0.85 cm³/g, preferably 0.30 to 0.75 cm³/g, preferably 0.35 to 0.70cm³/g, preferably 0.40 to 0.65 cm³/g, preferably 0.44 to 0.64 cm³/g. Insome embodiments, the porous silicate and/or aluminosilicate matrix isparticles of MCM-41 and the nanocarrier has a mean pore volume of 0.25to 0.65 cm³/g, preferably 0.30 to 0.60 cm³/g, preferably 0.35 to 0.55cm³/g, preferably 0.40 to 0.50 cm³/g, preferably 0.425 to 0.475 cm³/g,preferably 0.44 cm³/g. In some embodiments, the silicate and/oraluminosilicate matrix is particles of KIT-6 and the nanocarrier has amean pore volume of 0.45 to 0.85 cm³/g, 0.50 to 0.80 cm³/g, preferably0.55 to 0.75 cm³/g, preferably 0.60 to 0.70 cm³/g, preferably 0.625 to0.675 cm³/g, preferably 0.64 cm³/g.

In general, the antioxidant may be any suitable antioxidant known to oneof ordinary skill in the art. Examples of such antioxidants include, butare not limited to curcumin (and curcumin derivatives known ascurcuminoids), Coenzyme Q10, quercetin, rutin, ascorbic acid, gallicacid, edaravone, N-acetylcysteine, alfa-lipoic acid, diosmin,hesperidin, oxerutins, baicalein, tocotrienols, resveratrol or otherstilbenoids such as pterostilbene, retinoids and carotenes includingVitamin A, beta carotene, and alpha-carotene, astaxanthin,canthaxanthin, lutein, lycopene, and zeaxanthin, natural phenolsincluding flavonoids, silymarin, xanthones, eugenol, phenolic acids,lipoic acid, acetylcysteine, uric acid, glutathione, and catechin. Insome embodiments, the antioxidant is at least one selected from thegroup consisting of quercetin, rutin, coenzyme Q10, gallic acid, andcurcumin.

Quercetin has the following chemical structure:

Quercetin is a plant flavonol from the flavonoid group. It is found in awide variety of food sources, but has very low water solubility andbioavailability. Inclusion of quercetin in the nanomedicinal compositionof the present invention may overcome these disadvantageous propertiesof quercetin to increase an amount of quercetin which is delivered.Quercetin may be present in a crystalline or amorphous form or in amixture of both crystalline and amorphous forms, for example at a ratioof 1-99 wt. %:99-1 wt. %, 10-90 wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt.%, 30-70 wt. %:70-30 wt. % , 40-60 wt. %:60-40 wt. % or about 50 wt.%:about 50 wt. % (or any intermediate ratio of crystalline: amorphousforms).

Rutin has the following structure:

Rutin is the glycoside combining the flavonol quercetin and thedisaccharide rutinose (α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranose).Rutin may be present in a crystalline or amorphous form or in a mixtureof both crystalline and amorphous forms, for example at a ratio of 1-99wt. %:99-1 wt. %, 10-90 wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt. %,30-70 wt. %:70-30 wt. %, 40-60 wt. %:60-40 wt. % or about 50 wt. %:about50 wt. % (or any intermediate ratio of crystalline:amorphous forms).

Coenzyme Q10 (CoQ10) conforms to the following chemical structure:

CoQ10 is a 1,4-benzoquinone, where Q refers to the quinone chemicalgroup and 10 refers to the number of isoprenyl chemical subunits in itstail. Other forms of Coenzyme Q may be distinguished from CoQ10 by theirnumber of isoprenyl subunits. A CoQ such as CoQ10 may be present in acrystalline or amorphous form or in a mixture of both crystalline andamorphous forms, for example at a ratio of 1-99 wt. %:99-1 wt. %, 10-90wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt. %, 30-70 wt. %:70-30 wt. % ,40-60 wt. %:60-40 wt. % or about 50 wt. %:about 50 wt. % (or anyintermediate ratio of crystalline:amorphous forms).

Gallic acid has the following structure:

Gallic acid is a potent antioxidant against cancers (leukemia, colon andlung cancer cells) and other metabolic disorders. Gallic acid may bepresent in a crystalline or amorphous form or in a mixture of bothcrystalline and amorphous forms, for example at a ratio of 1-99 wt.%:99-1 wt. %, 10-90 wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt. %, 30-70wt. %:70-30 wt. %, 40-60 wt. %:60-40 wt. % or about 50 wt. %:about 50wt. % (or any intermediate ratio of crystalline:amorphous forms).

Curcumin has the following structure:

A curcuminoid is a linear diarylheptanoid. This class of compoundsincludes curcumin in both its keto and enolate forms as well as curcuminderivatives such as demethoxycurcumin and bisdemethoxycurcumin and theirgeomentrical isomers and metabolites including sulfate conjugates andglucoronides. Other examples of curcumin derivatives or analogs includethose described by Raja, et al., U.S. Pat. No. 9,447,023 B2, Raja, etal., U.S. Pat. No. 9,650,404 B2, Johnson, et al., U.S. Pat. No.9,556,105 B2 or Vander Jagt, et al., U.S. Pat. No. 9,187,397 B2 (allincorporated by reference); especially for their descriptions ofcurcuminoid formulas and various chemical species of curcuminoids. Insome embodiments of the invention curcumin or another curcuminoid may beincluded as an antioxidant in the nanomedicinal composition of thepresent disclosure.

Mixtures of curcuminoids are also contemplated such as one isolated fromrhizomes of turmeric comprised of Curcumin (75-81%), Demethoxycurcumin(15-19%) and Bisdemethoxycurcumin (2.5-6.5%). The content of any one ofa curcuminoid in a mixture may range from about 0 to about 100 wt. %,for example, 10-90 wt. %, 20-80 wt. %, 30-70 wt. %, 40-60wt %, 50 wt. %,40 wt. %, 33.3 wt. %, 30 wt. %, 20 wt. %, 10 wt. % or 5wt % or 1 wt. %.A mixture may contain two, three or more different curcuminoids.

Curcumin may be present in a crystalline or amorphous form or in amixture of both crystalline and amorphous forms, for example at a ratioof 1-99 wt. %:99-1 wt. %, 10-90 wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt.%, 30-70 wt. %:70-30 wt. % , 40-60 wt. %:60-40 wt. % or about 50 wt.%:about 50 wt. % (or any intermediate ratio of crystalline:amorphousforms). In some embodiments disclosed herein, curcumin will be in anamorphous form to increase its solubility.

Curcumin and its derivatives are known for their antimicrobial,anti-oxidative, anti-inflammatory, and anti-cancer properties such asmalignancies in the brain or nervous system. Curcumin has also beenproposed as an agent to treat oxidative stress, such as oxidative stressin the brain, and for treatment of neurodegenerative disease likeAlzheimer' s disease (“AD”) or Parkinson's disease (“PD”); Lee, et al.,Curr. Neuropharmacol. 2013 July; 11(4):338-378 (incorporated byreference).

Curcumin may also be functionalized or prepared as a conjugate withanother moiety to modify or improve its pharmacokinetic properties. Forexample, curcumin can be adsorbed through functionalization to a silane,carboxylic acid, or biotin. Biocompatibility of acurcuminoid/hierarchical aluminosilicate can be increased by themodification with chitosan, or poly (D,L-lactide-co-glycolide), orpolyethylene glycol.

In some embodiments, the antioxidant is curcumin.

Resveratrol has the following structure:

Resveratrol is an antioxidant which belongs to the class of compoundsknown as stillbenoids, which are hydroxylated derivatives of stilbene(also known as 1,2-diphenylethene). The resveratrol may be resveratrolitself, or and a geomentrical isomer or metabolite, including sulfateconjugates and glucoronides, of resveratrol. Resveratrol may be presentin a crystalline or amorphous form or in a mixture of both crystallineand amorphous forms, for example at a ratio of 1-99 wt. %:99-1 wt. %,10-90 wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt. %, 30-70 wt. %:70-30 wt.% , 40-60 wt. %:60-40 wt. % or about 50 wt. %:about 50 wt. % (or anyintermediate ratio of crystalline:amorphous forms). The resveratrol maybe present as the trans isomer depicted above, a cis isomer, or in amixture of both the cis and trans isomers, for example at a ratio of1-99 wt. %:99-1 wt. %, 10-90 wt. %:90-10 wt. %; 20-80 wt. %:80-20 wt. %,30-70 wt. %:70-30 wt. % , 40-60 wt. %:60-40 wt. % or about 50 wt.%:about 50 wt. % (or any intermediate ratio of cis:trans forms).

In some embodiments, the antioxidant is resveratrol.

In some embodiments, the antioxidant is present in the nanomedicinalcomposition in an amount of 5 to 50 wt %, preferably 10 to 47.5 wt %,preferably 15 to 45 wt %, preferably 17.5 to 42.5 wt %, preferably 20 to40 wt %, preferably 22.5 to 37.5 wt %, preferably 25 to 35 wt %,preferably 26 to 34 wt %, preferably 27 to 33 wt %, preferably 28 to 31wt %, preferably 29 to 30 wt %, based on a total weight of nanomedicinalcomposition.

In some embodiments, the antioxidant or its constituent compounds mayinteract with the surface of the nanocarrier via any suitableinteraction known to one of ordinary skill in the art. Such interactionsmay be, for example physisorption (e.g. Van der Waals interactions),ion-ion interactions, ion-dipole interactions, dipole-dipoleinteractions, and hydrogen bonding. Such interaction may be through orinvolving appropriate functional groups on the antioxidant. Examples ofsuch functional groups include, but are not limited to oxygen-containingfunctional groups such as alcohols, alkoxides, carboxylic acids andcarboxylates, esters, ketones, and ethers; nitrogen-containingfunctional groups such as amines, amides, azides, diimides, imines,porphyrins, imides, isonitriles, nitriles, and nitro functional groups;phosphorous-containing functional groups such as phosphines, phosphites,phosphates, phosphonites, phosphonates, phosphinites, and phosphinates;and sulfur-containing functional groups such as thiols, thiolates,disulfides, sulfones, sulfonic acids and sulfonates, sulfoxides, thials,thioesters, thiosulfinates, thiocarboxylic acids and thiocarboxylates,sulfinic acids and sulfinates, thiocyanates, and isothiocyanates. Theantioxidant may be electrically neutral or may have a charge, the chargebeing either positive or negative. A compound which is electricallyneural may be devoid of charges or may have a combination of positiveand negative charges in equal number so as to balance to electricallyneutral (e.g. zwitterionic). A compound which is electrically neutralmay interact to an equal extent with or be disposed equally upon boththe interior and exterior surfaces of the nanocarrier. Alternatively, acompound which is electrically neutral may preferentially interact witheither the interior or exterior surface of the nanocarrier. A compoundwhich bears a positive charge may preferentially interact with or bedisposed upon the exterior surface of the nanocarrier which bears anegative charge. A compound which bears a negative charge maypreferentially interact with or be disposed upon the interior surface ofthe nanocarrier which bears a positive charge.

In some embodiments, the nanomedicinal composition comprises abiocompatible coating. Such a biocompatible coating may be disposed uponthe nanocarrier and/or the antioxidant. In general, the biocompatiblecoating may be any suitable coating known to one of ordinary skill inthe art. Examples of such suitable biocompatible coatings include, butare not limited to, agarose, agar, carrageen, alginic acid, alginate, analginic acid derivative, a hyaluronate derivative, a polyanionicpolysaccharide, chitin, chitosan, fibrin, a polyglycolide, apolylactide, a polycaprolactone, a dextran or copolymer thereof,polyvinyl pyrrolidone, a polyacrylate, a wax, apolyethylene-polyoxypropylene-block polymer, wool fat, poly(L-lacticacid), poly(DL-Lactic acid) copoly(lactic/glycolic acid), cellulose, acellulose derivative, a glycol, polylactide-polyglycolide,polymethyldisiloxane, polycaprolactone, polylactic acid, and ethylenevinyl acetate.

In some embodiments, the antioxidant is curcumin and the nanomedicinalcomposition releases greater than 20%, preferably greater than 20.5%,preferably greater than 21%, preferably greater than 21.5%, preferablygreater than 22%, preferably greater than 22.5%, preferably greater than23%, preferably greater than 23.5%, preferably greater than 24%,preferably greater than 24.5%, preferably greater than 25% of a totalweight of curcumin within 24 to 72 hours, preferably 27 to 69 hours,preferably 30 to 66 hours, preferably 33 to 63 hours, preferably 36 to60 hours, preferably 39 to 57 hours, preferably 42 to 54 hours ofcontact with a suitable biological medium. In some embodiments,antioxidant is resveratrol the nanomedicinal composition releasesgreater than 7.5%, preferably greater than 8%, preferably greater than8.5%, preferably greater than 9%, preferably greater than 9.5%,preferably greater than 10%, preferably greater than 10.5%, preferablygreater than 11%, preferably greater than 11.5%, preferably greater than12%, preferably greater than 12.5% of a total weight of resveratrolwithin 24 to 72 hours, preferably 27 to 69 hours, preferably 30 to 66hours, preferably 33 to 63 hours, preferably 36 to 60 hours, preferably39 to 57 hours, preferably 42 to 54 hours of contact with a suitablebiological medium. Examples of suitable biological media include, butare not limited to, buffered saline solutions such as phosphate bufferedsaline, cell culture media such as Minimum Essential Medium (MEM, alsoknown as Eagle's minimal essential medium EMEM), Dulbecco's ModifiedEagle's Medium (DMEM), Iscove's Modified Dulbecco's Medium (IMDM),RPMI-1640, Ham's F-10, and F-12; animal tissue, or a subject's body.

In this aspect of the invention the release of the antioxidant may bedue over a release period of at least 2 hours, preferably at least 4hours, preferably at least 6 hours, preferably at least 8 hours,preferably at least 10 hours, preferably at least 12 hours, preferablyat least 14 hours, preferably at least 16 hours, preferably at least 18hours, preferably at least 20 hours Initial release rates are preferably10-20 wt % of the total amount of antioxidant in the nanomedicinalcomposition during the induction period. Upon passage of the inductionperiod and arrival of the nanomedicinal composition at a target site, amajor portion of the antioxidant is released. In some embodiments, themajor portion comprises at least 25 wt %, preferably at least 30 wt %,preferably at least 35 wt %, preferably at least 40 wt %, preferably atleast 45 wt %, preferably at least 50 wt % of a total amount ofantioxidant released.

In some embodiments, the induction period is provided by a coatingdisposed on the nanomedicinal composition, the coating as describedabove. In such embodiments, the coating may inhibit the release of theantioxidant. Removal of the coating by any suitable process, for exampleby dissolving, degrading, or digesting, may allow the pharmaceuticalgent mixture to be released.

In some embodiments, the nanomedicinal composition has an antioxidantrelease rate of 0.1 to 10 wt % per hour, preferably 0.25 to 9 wt % perhour, preferably 0.5 to 7.5 wt % per hour, preferably 0.75 to 5 wt % perhour, preferably 1 to 4 wt % per hour based on a total initial weight ofantioxidant. In such embodiments, the antioxidant release rate may be anaverage antioxidant release rate measured over the release period asdescribed above. In some embodiments, the nanomedicinal composition hasan initial antioxidant release rate which is maintained over an initialrelease period. In such embodiments, the initial release period may befollowed by a second release period which has a second antioxidantrelease rate. The initial antioxidant release rate and/or secondantioxidant release rate may be average release rates as describedabove. In some embodiments, the initial release period comprises thefirst hours of release, preferably the first 18 hours of release,preferably the first 16 hours of release, preferably the first 14 hoursof release, preferably the first 12 hours of release, preferably thefirst 10 hours of release. Such “first hours of release” are preferablymeasured from the initiation of release. The initiation of release maybe measured by, for example a delivery of the nanomedicinal compositionto a tumor site or a pre-determined amount of time after administration.Such a pre-determined time may be any suitable amount of time known toone of ordinary skill in the art, for example, an expected time fordelivery of the nanomedicinal composition to the tumor site, an expectedcirculation time, an expected coating degradation time, or the like.

In some embodiments, the nanomedicinal composition reduces the viabilityof Blastocystis organisms by at least 75%, preferably by at least 77.5%,preferably at least 80%, preferably at least 82.5%, preferably at least85%, preferably at least 87.5%, preferably at least 90%, preferably atleast 92.5%, preferably at least 95%, preferably at least 97.5%,preferably at least 99% when the nanomedicinal composition is presentfor 24 hours in an amount of 100 to 1000 μg/mL.

The present disclosure also relates to a method of forming thenanomedicinal composition, the method comprising mixing the metalorganic framework and the porous silicate and/or aluminosilicate matrixto form the nanocarrier, combining the nanocarrier and the antioxidantin an impregnation solution thereby forming the nanomedicinalcomposition, and isolating the nanomedicinal composition.

In general, the metal organic framework and the porous silicate and/oraluminosilicate matrix may be mixed using any suitable technique orequipment known to one of ordinary skill in the art. The metal organicframework and the porous silicate and/or aluminosilicate matrix can bepresent as particles as described above. In some embodiments, the metalorganic framework and the porous silicate and/or aluminosilicate matrixare mixed as solids. That is, the mixing occurs between dry (i.e.without solvent) powders of the metal organic framework and the poroussilicate and/or aluminosilicate matrix. In some embodiments, the mixingoccurs as a slurry, a suspension, a dispersion, or other suitable wet(i.e. with solvent) form. In general, the solvent may be water, anaqueous solution, an organic solvent, or a mixture thereof. The aqueoussolution may comprise, for example, a surfactant, a coating material asdescribed above, a surface functionalization material as describedabove, a salt, or mixtures thereof. The organic solvent may be anysuitable organic solvent, examples of which include, but are not limitedto alcohols such as methanol, ethanol, n-propanol, 2-propanol (alsoknown as isopropanol), ethylene glycol, diethylene glycol, and glycerol;hydrocarbons such as pentane, hexane, and heptane; ketones such asacetone and methyl ethyl ketone; esters such as ethyl acetate; amidessuch as dimethylformamide; ethers such as tetrahydrofuran, diglyme, anddiethyl ether; nitriles such as acetonitrile; halogenated organicsolvents such as methylene chloride (also known as dichloromethane),carbon tetrachloride, and chloroform; aromatic organic solvents such asbenzene and xylene; amines such as trimethylamine and pyridine; andmixtures thereof.

In some embodiments, the mixing comprises ultrasonication. Suchultrasonication may occur in a dry powder as described above or in aslurry, a suspension, a dispersion, or other suitable wet form asdescribed above.

In some embodiments, the impregnation solution comprises an alcoholhaving 1 to 5 carbon atoms. In some embodiments, the alcohol having 1 to5 carbon atoms is methanol. In some embodiments, the impregnationsolution comprises water. In some embodiments, the impregnation solutioncomprises glycerol. In some embodiments, the antioxidant is present inan amount of 0.5 to 3 mg/mL, preferably 0.75 to 2. 5 mg/mL, preferably1.0 to 2.25 mg/mL preferably 1.25 to 2.0 mg/mL, preferably 1.5 to 1.75mg/mL, preferably 1.6 to 1.7 mg/mL of impregnation solution.

The present disclosure also relates to a method for method for treatingan infection by a parasite in the genus Blastocystis in a subject, themethod comprising administering to a subject in need of therapy apharmaceutical composition comprising the nanomedicinal composition.Blastocystis is a genus of single-celled heterokont parasites, somespecies of which are capable of causing infection in a variety ofanimals, including mammals, birds, rodents, reptiles, amphibians, fish,and insects. Typically, infections occur in the gastrointestinal tract.Blastocystis has various morphological forms. Four commonly describedforms are the vacuolar (otherwise known as central body), granular,amoeboid, and cyst forms. An infection treated by the present inventionmay involve any of these forms. Infection with a Blastocystis parasitecan be referred to as “Blastocystosis”.

The pharmaceutical composition may further comprise one or morepharmaceutically acceptable excipients.

As used herein, a “pharmaceutically acceptable excipient” refers to acarrier, adjuvant, vehicle, diluent, or other inert substance useful tofurther facilitate administration of a compound that does not causesignificant irritation to an organism, does not abrogate the biologicalactivity and properties of the administered active ingredient, and/ordoes not interact in a deleterious manner with the other components ofthe composition in which it contains. The term “carrier” encompasses anyexcipient, binder, diluent, filler, salt, buffer, solubilizer, lipid,stabilizer, or other material well-known in the art for use inpharmaceutical formulations. The choice of a carrier for use in apharmaceutical composition will depend upon the intended route ofadministration for the pharmaceutical composition. The preparation ofpharmaceutically acceptable carriers and formulations containing thesematerials is described in, e.g. Remington's Pharmaceutical Sciences,21st Edition, ed. University of the Sciences in Philadelphia,Lippincott, Williams & Wilkins, Philadelphia Pa., 2005, which isincorporated herein by reference in its entirety). Examples ofphysiologically acceptable carriers include antioxidants includingascorbic acid; low molecular weight (less than about 10 residues)peptides; proteins, such as serum albumin, gelatine, or immunoglobulins;hydrophilic polymers such as polyvinylpyrrolidone; amino acids such asglycine, glutamine, asparagine, arginine or lysine; monosaccharides,disaccharides, and other carbohydrates including glucose, mannose, ordextrin; chelating agents such as EDTA; sugar alcohols such as mannitolor sorbitol; salt-forming counter ions such as sodium; and/or non-ionicsurfactants such as TWEEN® (ICI, Inc.; Bridgewater, N.J.), polyethyleneglycol (PEG), and PLURONICS™ (BASF; Florham Park, N.J.). Examples ofother inert substance added to a pharmaceutical composition to furtherfacilitate administration of a compound, without limitation, includecalcium carbonate, calcium phosphate, various sugars and types ofstarch, cellulose derivatives, gelatine, vegetable oils, andpolyethylene glycols.

In some embodiments, the pharmaceutically acceptable excipient is atleast one selected from the group consisting of a buffer, an inorganicsalt, a fatty acid, a vegetable oil, a synthetic fatty ester, asurfactant, and a polymer.

Exemplary buffers include, without limitation, phosphate buffers,citrate buffer, acetate buffers, borate buffers, carbonate buffers,bicarbonate buffers, and buffers with other organic acids and salts.

Exemplary inorganic salts include, without limitation, calciumcarbonate, calcium phosphate, disodium hydrogen phosphate, potassiumhydrogen phosphate, sodium chloride, zinc oxide, zinc sulfate, andmagnesium trisilicate.

Exemplary fatty acids include, without limitation, an omega-3 fatty acid(e.g., linolenic acid, docosahexaenoic acid, eicosapentaenoic acid) andan omega-6 fatty acid (e.g., linoleic acid, eicosadienoic acid,arachidonic acid). Other fatty acids, such as oleic acid, palmitoleicacid, palmitic acid, stearic acid, and myristic acid, may be included.

Exemplary vegetable oils include, without limitation, avocado oil, oliveoil, palm oil, coconut oil, rapeseed oil, soybean oil, corn oil,sunflower oil, cottonseed oil, and peanut oil, grape seed oil, hazelnutoil, linseed oil, rice bran oil, safflower oil, sesame oil, brazil nutoil, carapa oil, passion fruit oil, and cocoa butter.

Exemplary synthetic fatty esters include, without limitation, methyl,ethyl, isopropyl and butyl esters of fatty acids (e.g., isopropylpalmitate, glyceryl stearate, ethyl oleate, isopropyl myristate,isopropyl isostearate, diisopropyl sebacate, ethyl stearate, di-n-butyladipate, dipropylene glycol pelargonate), C12-C16 fatty alcohol lactates(e.g., cetyl lactate and lauryl lactate), propylene dipelargonate,2-ethylhexyl isononoate, 2-ethylhexyl stearate, isopropyl lanolate,2-ethylhexyl salicylate, cetyl myristate, oleyl myristate, oleylstearate, oleyl oleate, hexyl laurate, isohexyl laurate, propyleneglycol fatty ester, and polyoxyethylene sorbitan fatty ester. As usedherein, the term “propylene glycol fatty ester” refers to a monoether ordiester, or mixtures thereof, formed between propylene glycol orpolypropylene glycol and a fatty acid. The term “polyoxyethylenesorbitan fatty ester” denotes oleate esters of sorbitol and itsanhydrides, typically copolymerized with ethylene oxide.

Surfactants may act as detergents, wetting agents, emulsifiers, foamingagents, and dispersants. Surfactants that may be present in thecompositions of the present disclosure include zwitterionic (amphoteric)surfactants, e.g., phosphatidylcholine, and3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS),anionic surfactants, e.g., sodium lauryl sulfate, sodium octanesulfonate, sodium decane sulfonate, and sodium dodecane sulfonate,non-ionic surfactants, e.g., sorbitan monolaurate, sorbitanmonopalmitate, sorbitan trioleate, polysorbates such as polysorbate 20(Tween 20), polysorbate 60 (Tween 60), and polysorbate 80 (Tween 80),cationic surfactants, e.g., decyltrimethylammonium bromide,dodecyltrimethyl-iammonium bromide, tetradecyltrimethylammonium bromide,tetradecyltrimethylammonium chloride, and dodecylammonium chloride, andcombinations thereof.

Exemplary polymers include, without limitation, polylactides,polyglycolides, polycaprolactones, polyanhydrides, polyurethanes,polyesteramides, polyorthoesters, polydioxanones, polyacetals,polyketals, polycarbonates, polyorthocarbonates, polyphos-phazenes,polyhydroxybutyrates, polyhydroxyvalerates, polyalkylene oxalates,polyalkylene succinates, poly(malic acid), poly(maleic anhydride), apolyvinyl alcohols, and copolymers, terpolymers, or combinations ormixtures therein. The copolymer/terpolymer may be a randomcopolymer/terpolymer, or a block copolymer/terpolymer.

Depending on the route of administration e.g. oral, parental, ortopical, the pharmaceutical composition may be in the form of soliddosage form such as tablets, caplets, capsules, powders, and granules,semi-solid dosage form such as gels, pastes, and suppositories, liquiddosage forms such as suspension, and dispersions, inhalation dosage formsuch as aerosols, sprays, and powders.

Solid dosage forms for oral administration can include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive ingredient is ordinarily combined with one or more adjuvantsappropriate to the indicated route of administration. If administeredper os, the active ingredient can be admixed with lactose, sucrose,starch powder, cellulose esters of alkanoic acids, cellulose alkylesters, talc, stearic acid, magnesium stearate, magnesium oxide, sodiumand calcium salts of phosphoric and sulfuric acids, gelatine, acaciagum, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol,and then tableted or encapsulated for convenient administration. Suchcapsules or tablets can contain a controlled-release formulation as canbe provided in a dispersion of active compound in hydroxypropylmethylcellulose. In the case of capsules, tablets, and pills, the dosage formscan also comprise buffering ingredients such as sodium citrate,magnesium or calcium carbonate or bicarbonate. Tablets and pills canadditionally be prepared with enteric coatings.

Liquid dosage forms for oral administration can include pharmaceuticallyacceptable emulsions, suspensions, syrups, and elixirs containing inertdiluents commonly used in the art, such as water. Such pharmaceuticalcompositions can also comprise adjuvants, such as wetting ingredients,emulsifying and suspending ingredients, and sweetening, flavoring, andperfuming ingredients.

For therapeutic purposes, formulations for parenteral administration canbe in the form of aqueous or non-aqueous isotonic sterile injectiondispersions or suspensions. The term “parenteral”, as used herein,includes intravenous, intravesical, intraperitoneal, subcutaneous,intramuscular, intralesional, intracranial, intrapulmonal, intracardial,intrasternal, and sublingual injections, or infusion techniques. Thesedispersions and suspensions can be prepared from sterile powders orgranules having one or more of the carriers or diluents mentioned foruse in the formulations for oral administration. The active ingredientcan be dissolved in water, polyethylene glycol, propylene glycol,ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzylalcohol, sodium chloride, and/or various buffers. Other adjuvants andmodes of administration are well and widely known in the pharmaceuticalart.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting ingredients and suspendingingredients. The sterile injectable preparation can also be a sterileinjectable dispersion or suspension in a non-toxic parenterallyacceptable diluent or solvent, for example, in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil can be employedincluding synthetic mono- or di-glycerides. In addition, fatty acids,such as oleic acid, find use in the preparation of injectable.Dimethylacetamide, surfactants including ionic and non-ionic detergents,polyethylene glycols can be used. Mixtures of solvents and wettingingredients such as those discussed above are also useful.

Suppositories for rectal administration can be prepared by mixing theactive ingredient with a suitable non-irritating excipient, such ascocoa butter, synthetic mono-, di-, or triglycerides, fatty acids, andpolyethylene glycols that are solid at ordinary temperatures but liquidat the rectal temperature and will therefore melt in the rectum andrelease the drug. Such suppositories may be advantageous for treatingcolorectal infections, but may be unsuitable for treating otherinfection locations.

Administration by inhalation may be advantageous for treating lunginfections, but may be unsuitable for treating other infectionlocations.

In other embodiments, the pharmaceutical composition comprising thenanomedicinal composition disclosed herein thereof has different releaserates categorized as immediate release and controlled- orsustained-release.

As used herein, the terms “treat”, “treatment”, and “treating” in thecontext of the administration of a therapy to a subject in need thereofrefers to the reduction or inhibition of the progression and/or durationof a disease (e.g. infection with a parasite in the genus Blastocystis),the reduction or amelioration of the severity of the disease, and/or theamelioration of one or more symptoms thereof resulting from theadministration of one or more therapies. “Treating” or “treatment” ofthe disease includes preventing the disease from occurring in a subjectthat may be predisposed to the disease but does not yet experience orexhibit symptoms of the disease (prophylactic treatment), inhibiting thedisease (slowing or arresting its development), ameliorating thedisease, providing relief from the symptoms or side-effects of thedisease (including palliative treatment), and relieving the disease(causing regression of the disease). With regard to the disease, theseterms simply mean that one or more of the symptoms of the disease willbe reduced. Such terms may refer to one, two, three, or more resultsfollowing the administration of one, two, three, or more therapies: (1)a stabilization, reduction (e.g. by more than 10%, 20%, 30%, 40%, 50%,preferably by more than 60% of the population of parasite in the genusBlastocystis cells before administration), or elimination of theparasite in the genus Blastocystis cells, (2) inhibiting parasite in thegenus Blastocystis cell division and/or parasite in the genusBlastocystis cell proliferation, (3) relieving to some extent (or,preferably, eliminating) one or more symptoms associated with apathology related to or caused in part by infection with a parasite inthe genus Blastocystis, (4) an increase in disease-free, relapse-free,progression-free, and/or overall survival, duration, or rate, (5) adecrease in hospitalization rate, (6) a decrease in hospitalizationlength, (7) eradication, removal, or control of primary, regional and/ormetastatic parasite in the genus Blastocystis, (8) a stabilization orreduction (e.g. by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%,preferably at least 80% relative to the initial growth rate) in thegrowth of cells of a parasite in the genus Blastocystis, (9) a reductionin mortality, (10) an increase in the response rate, the durability ofresponse, or number of patients who respond or are in remission, (11) adecrease in the need for treatment by another therapeutic, and (12)preventing or reducing (e.g. by more than 10%, more than 30%, preferablyby more than 60% of the population of cells of parasite in the genusBlastocystis.

The term “subject” and “patient” are used interchangeably. As usedherein, they refer to any subject for whom or which therapy, includingwith the pharmaceutical compositions according to the present disclosureis desired. In most embodiments, the subject is a mammal, including butis not limited to a human, a non-human primate such as a chimpanzee, adomestic livestock such as a cattle, a horse, a swine, a pet animal suchas a dog, a cat, and a rabbit, and a laboratory subject such as arodent, e.g. a rat, a mouse, and a guinea pig. In preferred embodiments,the subject is a human.

The terms “administer”, “administering”, “administration”, and the like,as used herein, refer to the methods that may be used to enable deliveryof the active ingredient and/or the composition to the desired site ofbiological action. Routes or modes of administration are as set forthherein. These methods include, but are not limited to, oral routes,intraduodenal routes, parenteral injection (including intravenous,subcutaneous, intraperitoneal, intramuscular, intravascular, orinfusion), and rectal administration. Those of ordinary skill in the artare familiar with administration techniques that can be employed withthe compounds and methods described herein. In preferred embodiments,the active ingredient and/or the pharmaceutical composition describedherein are administered orally.

The dosage amount and treatment duration are dependent on factors, suchas bioavailability of a drug, administration mode, toxicity of a drug,gender, age, lifestyle, body weight, the use of other drugs and dietarysupplements, the infection stage or severity, tolerance and resistanceof the body to the administered drug, etc., and then determined andadjusted accordingly. The terms “effective amount”, “therapeuticallyeffective amount”, “pharmaceutically effective amount” or “sufficientamount” refer to that amount of the active ingredient being administeredwhich will relieve to some extent one or more of the symptoms of theinfection or disease being treated. The result can be a reduction and/oralleviation of the signs, symptoms, or causes of a disease, or any otherdesired alteration of a biological system. An appropriate “effectiveamount” may differ from one individual to another. An appropriate“effective amount” in any individual case may be determined usingtechniques, such as a dose escalation study. In some embodiments, aneffective amount is in the range of 0.1-30 g/kg of the nanomedicinalcomposition per body weight of the subject.

A treatment method may comprise administering the pharmaceuticalcomposition of the current disclosure as a single dose or multipleindividual divided doses, wherein the nanomedicinal composition isaccumulated and releases the loaded antioxidant in or nearby theinfected tissues. In some embodiments, the pharmaceutical composition isadministered at various dosages (e.g. a first dose with an effectiveamount of nanomedicinal composition comprising 200 mg of the antioxidantper kilogram of the subject and a second dose with an effective amountof the nanomedicinal composition comprising 50 mg of the antioxidant perkilogram of the subject). In some embodiments, the interval of timebetween the administration of the pharmaceutical composition and theadministration of one or more additional therapies may be about 1-5minutes, 1-30 minutes, 30 minutes to 60 minutes, 1 hour, 1-2 hours, 2-6hours, 2-12 hours, 12-24 hours, 1-2 days, 2 days, 3 days, 4 days, 5days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 15 weeks, weeks, 26 weeks,52 weeks, 11-15 weeks, 15-20 weeks, 20-30 weeks, 30-40 weeks, 40-50weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2years, or any period of time in between. Preferably, the pharmaceuticalcomposition is administered once daily for at least 2 days, 5 days, 6days, or 7 days. In certain embodiments, the pharmaceutical compositionand one or more additional therapies are administered less than 1 day, 1week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 6 months,1 year, 2 years, or 5 years apart.

In some embodiments, the administration is stopped once the subject istreated.

The examples below are intended to further illustrate protocols forpreparing, characterizing, and using the nanomedicinal composition orfor treating an infection by a para

site in the genus Blastocystis using the nanomedicinal composition andare not intended to limit the scope of the claims.

Where a numerical limit or range is stated herein, the endpoints areincluded. Also, all values and subranges within a numerical limit orrange are specifically included as if explicitly written out.

Obviously, numerous modifications and variations of the presentinvention are possible in light of the above teachings. It is thereforeto be understood that, within the scope of the appended claims, theinvention may be practiced otherwise than as specifically describedherein.

EXAMPLES Nanocomposites

Bioactive naturally based polyphenols were nano-formulated, byaccommodating ZIF-8, structured nano-mesosilica (MCM-41 or KIT-6) andnatural antioxidants, curcumin or resveratrol. ZIF-8 with texturalproperties of 1040 m²/g (Z1200, sigma Aldrich), was purchased and usedas a composite with mesoporous silica. The hexagonal shaped nano-silica(Si-MCM-41) and three-dimensional Si-KIT-6 was synthesized usinghydrothermal technique. Cationic template cetyltrimethyl ammoniumbromide (CTAB) and non-ionic template (P123) were used respectively togenerate hexagonal and cubic pores for MCM-41 and KIT-6.

Synthesis of MCM-41 and KIT-6

For synthesis of MCM-41: 10.6 g of sodium metasilicate (silica source)was taken and dissolved in 50 ml of distilled water and stirredvigorously for 1 h. After dissolution, the solution was added dropwiseto 4.5 g of CTAB solution dissolved in 40 ml of distilled water. Themixture was further stirred for 2 h and then pH was adjusted to 10.5using diluted sulphuric acid solution (4N). The sol solution in milkyform was hydrothermally treated at 140° C. for 12-24 h.

For synthesis of KIT-6: 4g of P123 was dissolved in acidic HCl solution(2 M) and allowed to stir for 1 h. Then 4 g of n-butanol (co-solvent)was added along with 8.6g of tetraethylorthosilicate (silica source) andstirred for 24 h. The mixture in polypropylene bottle was transferred tooven and hydrothermally aged at 100° C. for 24 h. The precipitate wasfiltered, washed several times with excess water and dried at 100° C.for 12 h. The two samples (MCM-41 and KIT-6) were finally calcined at550° C. for 6 h.

Preparation of Medicinal Nanocomposite

The calcined form of MCM-41 and KIT-6 was composited with ZIF-8 usingultrasonic technique forming ZIF-8/MCM-41 and ZIF-8/KIT-6 nanocomposite(ZIF-8/structured silica ratio of 0.105). In order to nano-formulatewith bioactive polyphenol components, a rotary evaporator was used. Inbrief, curcumin or resveratrol (500 mg) were loaded over ZIF-8/MCM-41and ZIF-8/KIT-6 (2000 mg) in methanolic solution (300 ml) and sonicatedfor 10 min. Then the solvent was evaporated using rotary evaporatortechnique to form MCM-41/ZIF-8/antioxidant and KIT-6/ZIF-8/antioxidantnanocomposites. The formulated nanocomposites were characterized usingphase (XRD), textural features (BET), chemical coordination environmentof metal species (DRS-UV-visible), active component functionalization(FTIR) and transmission electron microscopy (JEM2100F from JEOL).

Drug Release Study

The release trend of antioxidants (resveratrol and curcumin) wereinvestigated using four different nanoformulationsMCM-41/ZIF-8/Curcumin, KIT-6/ZIF-8/Curcumin, MCM-41/ZIF-8/Resveratroland KIT-6/ZIF-8/Resveratrol nanocomposites. Prior to the study, thedialysis membrane (12 KDa, Sigma Aldrich) was activated and then 15 mgof nanoformulation was dispersed in 25 ml of PBS solution (pH 5.6 and7.4). The release of antioxidants are studied at 37° C. At regular timeinterval, 5 ml of solution was withdrawn, and release of antioxidantswere measured using UV-visible spectroscopy. The withdrawn solution (5ml) was replaced with equal volume of fresh PBS solution.

The release content was identified at specific wavelength of curcuminand resveratrol (307 nm). Prior to analysis, calibration curve forcurcumin and resveratrol were established. At first, an initial stocksolution was prepared with concentration of 1000 μg/ml for curcumin andresveratrol. Various concentration of aliquots 5, 10, 15, 20, 25 and 30μg/ml was prepared with makeup volume to 10 ml using release medium PBSsolution (pH=5.6 or 7.4) and calibration curve established against blankat maximum absorption wavelength λmax of 428 nm and 307 nm,respectively. Linear regression for curcumin and resveratrol were foundto be y=0.0041x+0.0237 and y=0.0579x+0.0318, where y corresponds toabsorbance and x to the concentration of antioxidant release (μg/ml).The correlation coefficient was of 0.9982 and 0.995 for maximumabsorption for curcumin and resveratrol, respectively. The release studywas repeated in triplicates.

Blastocystis Parasite

Suspensions of cultured Blastocystis from symptomatic patients were usedfor evaluating biological activities of the formulated nanocomposites.Blastocystis were obtained from the El-Badry Lab where ethicallyapproved by the Deanship of Scientific Research and Postgraduate Studiesof Imam Abdulrahman Bin Faisal University under the Institutional ReviewBoard (IRB) number (IRB #2021-01-009).

Blastocystis were sub-cultured using fresh Jones' medium enriched with10% horse serum according to Jones to get rid of stool debris [Jones, W.Ann Trop Med Parasitol., 1946, 130-140; & Zman, V., and Khan, K. SouthtAsian J Trop Med Public Health, 1994, 25, 792-793, each of which isincorporated herein by reference in its entirety]. Cultures withvacuolar forms of Blastocystis greater than 10⁶ were used for evaluatingbiological activities of the formulated nanocomposites.

Molecular Characterization of Isolated Blastocystis

One ml of subculture was suspended in 7 ml PBS, vortexed, centrifugedfor 1 min at 12.000×g, then the pellet was kept for DNA extraction. DNAwas extracted from pelleted Blastocystis cysts, using commercial DNAextraction kit (DNA MiniPrep™ kit, Zymo Research Corp., USA) followingthe kit's protocol. As per Stensvold's recommendations, Blastocystis DNAwas amplified and genotyped using two PCR reactions, targetingBlastocystis specific SSU rDNA and subtype-specific Sequence-Tagged-Site(STS) in order to detect the seven standardized subtypes (STs 1-7)[Stensvold, C. R. J Clin Microbiol., 2003, 51, 190-194, incorporatedherein by reference in its entirety]. PCR conditions and reactions wereperformed as mentioned by Yoshikawa et al. and Scicluna et al.[Yoshikawa, H., et. al., J Eukaryotic Microbiol., 2003, 50, 710-711; &Scicluna S M, et. al., Protist., 2006, 157, 77-85, each of which isincorporated herein by reference in its entirety].

In Vitro Challenging of Blastocystis With Nanocomposites

Suspension of cultured Blastocytis cysts, in logarithmic growth phase of10⁶/ml, were inoculated in sets of culture tubes with Jones' culturemedia. Haemocytometer counting chamber was used to count the number ofBlastocystis cysts. The experiments contained 3 groups as follows:

-   -   Group 1 (G1): parasite (negative, not treated) control group,        containing only cultured cysts of Blastocystis.    -   Group 2 (G2): drug (positive) control, containing cultured        parasites exposed to 500 μg/ml metronidazole as reference        anti-Blastocystis therapy.    -   Group 3 (G3): nanocomposites tested group, containing cultured        parasites exposed to the three nanocomposites. G3 was further        divided into subgroups to test three nanocomposites (G3A, G3B        and G2C) at different concentrations (G3i-iv). Parasites were        exposed to increasing concentrations of the three nanocomposites        of 100 μg/ml (G3i) 200 μg/ml (G3ii), 500 μg/ml (G3iii) and 800        μg/ml (G3iv) (W/V). All tubes were incubated at 37° C. in        humidified CO2 for 48 hours.

All tested nanocomposites for each concentration were prepared bydiluting stock solution in appropriate amount of PBS. The volume of eachof the experiment tubes was brought to 1 ml with Blastocystis 10⁶/mlconcentration. All experiments were performed in triplicate [El-Sayed,S. H., et. al., Res J Parasitol., 2017, 12,2, 33-44; Méabed, E. M. H.,Phytomedicine, 2018, 1, 43, 158-163; & Mokhtar, A. B., et. al., Int JEnviron Res Public Health, 2019, 16, 1555, each of which is incorporatedherein by reference in its entirety].

Assessment of Anti-Blastocystis Activities of the Nanocomposites

The number of Blastocystis cysts from all cultured tubes included in theexperiment were counted under the microscope in haemocytometer countingchamber, and cultured Blastocystis cysts were tested for their viabilityusing Trypan blue solution (0.4%). Trypan blue is a vital stain whichdoes not stain a live Blastocystis cysts, while it stain deadBlastocystis cysts blue. The dead Blastocystis cysts were furtherconfirmed by microscopic detection of cell wall disruption anddestruction of internal structures. All cultured tubes were examined forpercentage of reduction in growth of Blastocystis cysts each hour for 5hours then after 24 hours. The minimal lethal concentration (MLC) wasdetermined to be the concentration at which no Blastocystis cysts wereobserved.

Statistical Analysis

The obtained data was statistically analyzed using SPSS software andpresented as mean and standard deviation (SD). All the data shows themean for each of the three independent experiments. Means were comparedand variances were analyzed. P values of less than 0.05 indicatestatistical significance.

Results

MCM-41/ZIF-8/Curcumin, KIT-6/ZIF-8/Curcumin, MCM-41/ZIF-8/Resveratroland KIT-6/ZIF-8/Resveratrol and Zn nanocomposites were synthesized. TheXRD analysis of ZIF-8, Resveratrol, Curcumin, MCM-41/ZIF-8/Curcumin,KIT-6/ZIF-8/Curcumin, MCM-41/ZIF-8/Resveratrol andKIT-6/ZIF-8/Resveratrol nanocomposites are shown in FIG. 2 .

ZIF-8 exhibited typical crystalline peaks corresponding to sodalitestructure. Resveratrol and Curcumin indicated a characteristicscrystalline peaks. Four samples of composites were analyzed to study theextent of amorphous transformation of bioactive components in contactwith mesoporous silica. The characteristic peaks of ZIF-8 and curcuminwere not observed in XRD spectra of MCM-41/ZIF-8/Curcumin andKIT-6/ZIF-8/Curcumin. These findings indicate the amorphoustransformation of ZIF-8 and curcumin inside the hexagonal and cubicpores of MCM-41 and KIT-6, respectively. In case ofMCM-41/ZIF-8/Resveratrol, some peaks of resveratrol were seen, while incubic shaped KIT-6 none were observed. These results show that somecrystalline forms of resveratrol were still present inside the hexagonalpores of MCM-41. Overall, the transformation of ZIF-8 and antioxidantsfrom crystalline to amorphous form increased their bioavailability.

The textural properties of MCM-41, KIT-6, MCM-41/ZIF-8 and KIT-6/ZIF-8nanocomposites were measured using nitrogen adsorption technique (FIG.3A). The surface area, pore volume and average pore size values arepresented in Table 1. The adsorption-desorption isotherm pattern ofMCM-41 and KIT-6 samples exhibited a typical type IV isotherm withcapillary condensation due to large meso sized pores at 0.2-0.4 and0.6-0.8, respectively. The surface area of MCM-41 and KIT-6 was 942 m²/gand 897 m²/g, respectively. However, the formation of nanocomposite withZIF-8 reduced the surface area of MCM-41/ZIF-8 and KIT-6/ZIF-8 to 594m²/g and 336 m²/g. The similar trend in the pore volume and pore sizedistribution shows the effective interaction of structured silica withZIF-8 (FIG. 3B). The morphological analysis of KIT-6/ZIF-8 using TEMclearly shows the nanocomposite formation between KIT-6 and ZIF-8. Thesphere shaped ZIF-8 shown to be well distributed along with largesurface of KIT-6 (FIGS. 4A-4B).

TABLE 1 Textural and structural properties. BET surface Pore volumeAverage pore Sample area (m²/g) (cm³/g) size (nm) MCM-41 942 0.87 3.7KIT-6 897 1.24 5.5 MCM-41/ZIF-8 594 0.44 3.0 KIT-6/ZIF-8 336 0.64 7.6

FIG. 5A shows the FTIR spectra of Resveratrol, ZIF-8, MCM-41,MCM-41/ZIF-8/Resveratrol and KIT-6/ZIF-8/Resveratrol nanocomposites. Thespectra of resveratrol and ZIF-8 showed a characteristic bandcorresponding to carbon-carbon double bonds of aromatic compound at1605, 1583 and 1514 cm⁻¹. Phenolic compound containing a carbonyl groupshowed a band at 1155 cm⁻¹. The hydroxyl group from the phenoliccompound showed a stretching at 1390 cm⁻¹. The C—H group showed a bandat 960 cm⁻¹, indicating the trans resveratrol configuration. The C—Hvibration band of arene conjugated to olefinic group can be seen at 805cm⁻¹ and 836 cm⁻¹. In the various bands that are observed between650-500 cm⁻¹, a ═C—H of olefinic group can be seen at 670 cm⁻¹. Comparedto functional bands of hexagonal pores of MCM-41, various functionalpeaks of resveratrol in reduced signals were observed, indicating thepresence of amorphous components at the external pores, however, no suchpeaks of resveratrol were observed in the cubic shaped pores of KIT-6.This trend shows effective pore filling of nanosized resveratrol incubic pores of KIT-6. FIG. 5B shows the FTIR spectra of curcumin,MCM-41/ZIF-8/Curcumin and KIT-6/ZIF-8/Curcumin nanocomposites. Thefunctional groups of curcumin including >C═O and C═C were observedbetween 1627-1450 cm⁻¹. In particular, a characteristic enolic OH ofcurcumin can be observed at 962 cm⁻¹. The —C—O—C— chain vibrations(symmetric and asymmetric) can be observed between 1000-1450 cm⁻¹. Inthe case of both nanocomposites, the curcumin functional group showed areduction in >C═O and C═C peaks. A reduction in hydroxyl peak of enolgroup also indicates an overall effective interaction of curcuminthrough enol and other functional groups. In the case of MCM-41 andKIT-6, the broadening of peak at about 1023 cm⁻¹, indicates theeffective functionalization of curcumin in the hexagonal and cubic poresof nanocomposite.

FIG. 6 shows the diffuse reflectance spectra of KIT-6/ZIF-8,Resveratrol, Curcumin, MCM-41/ZIF-8/Curcumin, KIT-6/ZIF-8/Curcumin,MCM-41/ZIF-8/Resveratrol and KIT-6/ZIF-8/Resveratrol nanocomposites.KIT-6/ZIF-8 shows a strong absorption at 212 nm, which can be ascribedto the presence of Zn²⁺ species in ZIF-8. Resveratrol and curcuminrevealed broad absorption between 200-600 nm. After loading of curcuminand resveratrol, the absorption maximum increases significantly overMCM-41/ZIF-8 and KIT-6/ZIF-8/nanocomposites. Such expansion behaviorclearly indicates the composite formation over two supports.

Drug Delivery Study

The drug release ability of antioxidants on MCM-41/ZIF-8/Curcumin,KIT-6/ZIF-8/Curcumin, MCM-41/ZIF-8/Resveratrol andKIT-6/ZIF-8/Resveratrol nanocomposites was studied at intestinalparasite pH condition for human intestine ranging pH 5.3 to 7.4 (FIG. 5). Resveratrol (pure form) was studied for comparative purpose. In therelease study, as-such resveratrol in the absence of nanocarrier showeda burst release as expected and reached a maximum of 43% within 12 h. Incase of nanocomposite/resveratrol release profiles, MCM-41/ZIF-8exhibited a slow release of about 15% for 96 h, while KIT-6/ZIF-8 showedeven a lower release profile of about 7% for 96 h. At neutral pHcondition, both nanocomposites showed a reduced release of resveratrol.FIG. 5 shows the curcumin on KIT-6 support alone. A quick release ofcurcumin was observed reaching at about 37% within 3 h and then studiedand reached about 52% for 96 h. In case of nanocomposites, a widerdifference was observed in curcumin release. KIT-6/ZIF-8 showed a highpercentage of cumulative release of curcumin (39%) than MCM-41/ZIF-8(21%) at pH 5.6. MOFs are shown to exhibit pH sensitive drug releases.Further, the degradation of MOF at acidic, neutral and basic conditionstends to influence the drug release characteristics. MOF based onzirconium has shown to exhibit pH sensitive release of biphosphate baseddrug alendronate. The nanocarrier showed high drug loading efficiencyand pH sensitive release capability due to inherent anchoring nature ofmetal-oxygen clusters. The high release at acidic pH of 5.5 was mainlyattributed to the protonation of drug leading to high release thanneutral pH of 7.4 [Zhu X, et. al., Chemical Communications, 2014, 50,63, 8779-82; & Cai W, et. al., Advanced Science, 2019, 6, 1, 1801526,each of which is incorporated herein by reference in its entirety]. TheZIF-8 nanocarrier with pH sensitivity has been reported to be beneficialagainst parasite protozoan Trypanosoma infection. At acidic pH of 4.5, aquick release of benznidazole was observed, while the trend changes to asustained and longer release at pH 7.4 [de Moura Ferraz, L. R., et. al.,J Mater Sci: Mater Med, 2021, 32, 59, incorporated herein by referencein its entirety]. In the present nanocomposites, the hydroxylfunctionalization of curcumin and resveratrol with nanocomposites couldbe critical in protonation and assist such release in acidic pHcondition. Both MCM-41/ZIF-8 and KIT-6/ZIF-8 nanocomposites showed areduction in surface area and hierarchical pore formations compared toMCM-41 and KIT-6 (FIG. 7 ). In parallel, an intermediate pH sensitiverelease with steady modulation of antioxidants occurs in KIT-6/ZIF-8than KIT-6 and resveratrol.

Anti-Blastocystis Activity

The anti-Blastoystis cysts activities of the nanocomposites, weredetermined by calculating the viability percent of Blastoystis cystsexposed to different increasing concentrations of all the nanocompositeseach hour for 5 hours then after 24 hours (100, 200, 500 and 800 μg/l).The parasite control group showed no effect on viability of Blastoystiscysts. There was a reduction in number of cysts exposed to thenanocomposite; the maximum effect was observed withResveratrol/ZIF-8/MCM-41 (see Table 2).

Resveratrol/ZIF-8/MCM-41 showed the maximum percentage of Blastocystiscysts destruction (˜100%) at concentration of 500 μg/ml after 5 hours ofexposure as found using standard drug (metronidazole) with statisticalsignificance.

All of the Curcumin/ZIF-8/MCM-41 nanocomposites tested concentrationsand three concentrations (200, 500 and 800 μg/ml) of theantioxidant-free nanocomposite killed more than 90% of the viableBlastocystis cysts in one day compared to parasite control, untreatedBlastocystis cysts. The percentage of viability of Blastoystis cystssteadily declined over the experiment time; it was dose-dependent in alltested nanocomposites, with stronger effect using resveratrol, which wasable to kill >90 of Blastocystis cysts in one hour (Table 2). There wasa statistical significance in low doses of the nanocomposites (Table 2).The plots of these results are shown in FIGS. 8A-8C.

TABLE 2 Reduction in the percentage of viability of Blastocystis cystsin parasite control group (negative control), after exposure tometronidazole (positive control), and after exposure to the threenanocomposites. Time of exposure/hour 1 hour 2 hours 3 hours 4 hours 5hours 24 hours Parasite control  0.67 ±  0.67 ±  0.33 ±  0.67 ±  0.67 ± 1.67 ± 0.58 1.15 0.58 0.58 1.15 0.58 Metronidazole control 81.67 ±87.67 ± 91.33 ± 96.33 ± 98.67 ± 99.33 ± 2.52 1.53 1.53 1.15 0.58 0.58Resvetrol/ 100 Mean ± 50.33 ± 56.67 ± 61.67 ± 70.33 ± 84.67 ± 91.33 ±ZIF8/ μg/ml Sd 3.06 2.52 3.06 2.08 2.31 4.16 KIT-6 P. value    0.0004*   0.0002*    0.0005*    0.0004*    0.0003*    0.0003* 200 Mean ± 57.33± 63.67 ± 72.33 ± 80.33 ± 92.33 ± 95.67 ± μg/ml Sd 5.51 4.04 4.16 3.064.04 4.16 P. value   0.001*   0.001*    0.0007*    0.0003*    0.0003*  0.004* 500 Mean ± 76.33 ± 84.33 ± 91.33 ± 96.67 ± 99.33 ± 99.67 ±μg/ml Sd 4.04 6.11 5.68 2.08 0.58 0.58 P. value    0.0004*    0.0006*   0.0008*    0.0001*  0.04*  0.05* 800 Mean ± 90.67 ± 94.33 ± 97.33 ±99.33 ± 99.67 ± 99.67 ± μg/ml Sd 2.08 4.73 2.89 0.58 0.58 0.58 P. value  0.008*    0.0003*    0.0001* 0.21 0.42 0.12 Curcumin/ 100 Mean ± 39.67± 43.34 ± 48.67 ± 53.34 ± 57.34 ± 74.67 ± ZIF-8/ μg/ml Sd 2.3  3.05 6.033.21 2.08 3.51 MCM-41 P. value    0.0004*    0.0002*    0.0005*   0.0004*    0.0003*    0.0003* 200 Mean ± 44.34 ± 50.34 ± 54.34 ±75.34 ± 84.34 ± 90.67 ± μg/ml Sd 2.08 2.52 3.06 2.08 3.51 4.73 P. value  0.001*   0.001*    0.0007*    0.0003*    0.0003*   0.001* 500 Mean ±68.34 ± 73.34 ± 78.34 ± 86.67 ± 91.33 ± 95.34 ± μg/ml Sd 3.79 3.06 2.522.52 2.08 2.08 P. value    0.0004*    0.0006*    0.0008*    0.0001* 0.04   0.0001* 800 Mean ± 85.00 ± 87.67 ± 90.34 ± 93.33 ± 96.67 ± 99.67 ±μg/ml Sd 2.65 2.08 2.08 2.89 0.58 0.58 P. value   0.008*    0.0003*   0.0001* 0.21 0.42    0.0001* ZIF-8/ 100 Mean ± 36.33 ± 40.33 ± 44.67± 49.33 ± 56.67 ± 63.33 ± KIT-6 μg/ml Sd 0.58 1.15 1.15 0.58 0.58 1.15P. value    0.0003*    0.0006*    0.0008*   0.001*   0.005*  0.035 200Mean ± 41.67 ± 45.67 ± 51.33 ± 56.33 ± 62.67 ± 75.67 ± μg/ml Sd 1.531.53 0.58 0.58 0.58 1.53 P. value   0.003*   0.002*   0.003*   0.0045 0.034  0.135 500 Mean ± 49.33 ± 54.67 ± 58.67 ± 64.67 ± 78.33 ± 85.33 ±μg/ml Sd 0.58 1.15 1.53 1.53 2.08 0.58 P. value   0.005*   0.021*  0.053*   0.047*   0.033*  0.211 800 Mean ± 72.67 ± 76.33 ± 77.67 ±81.67 ± 86.67 ± 91.33 ± μg/ml Sd 1.15 3.06 2.52 2.52 2.08 0.58 P. value  0.008*  0.100  0.021  0.072 0.15 0.40 Data were presented as the mean± standard deviation (SD), *P < 0.05 is statistically significant,compared to the parasite control.

To date, there is no full cure for Blastocystosis, an emerging parasiticdisease. Small doses of metronidazole (as of 10 ug/ml) have been used asin-vitro anti-Blastocystis therapy, however, higher doses up to 1 mg/mlare typically required [Haresh, K., et. al., Isolate resistance ofBlastocystis hominis to metronidazole., Trop Med Int Hlth., 1999, 4,274-277, incorporated herein by reference in its entirety]. Here, ahigher dose of 500 ug/ml of metronidazole was used as a comparison.

Resveratrol showed promising anti-protozoal activity, but itsanti-Blastocystis activity has not yet been evaluated. Here, the invitro effects of resveratrol and ZIF-8/KIT-6 nanocomponents on theviability of Blastocystis cysts were assessed. There was dose-dependent,and time depended anti-Blastocystis cysts activity for the two assessednanocomposites at a low concentration. The MLC of resveratrolnanocomponents was 100 ug/ml, which is a very small dose than the higherdoses tested before. Juan et al. reported that the prolonged eating (for28 days) of large doses of resveratrol (1.4 gm), 1000 times the amountingested daily by a human of 70 kg, causes no harm to rat as compared torat control group. There were no histological, biochemical, orhematological changes in treated rats, also there was no change inhabits of drinking water or consumption of food of treated rats, norchange of body weight [Juan, M.E., et. al., J Nutr., 2002, 132, 257-260,incorporated herein by reference in its entirety].

Studies have attributed the anti-protozoal activities of resveratrol andcurcumin to reduced oxygen consumption by protozoa, which may explainits mechanism of action as anti-Blastocystis [Leiro, J., et. al.,Antimicrob Agents Chemother., 2004, 48, 2497-2501; Kedzierski, L., et.al., Parasitol Res., 2007, 102, 91-97; Lamas, J., et. al., VetParasitol., 2009, 161, 307-315; & Vang, O., et. al., PLoS One, 2011, 6,e19881, each of which is incorporated herein by reference in itsentirety]. There are many advantages to the nano-scaling of resveratroland curcumin. It enhances their therapeutic activities by inducingcellular stress responses at lower doses. This low dose adaptiveresponse is due to its nano-size with large surface area to volumeratio. Giving the drug in lower doses reduces toxicity. In addition,with prolonged exposure, all nanocomposites killed Blastocystis cysts,this may be because nanocomposites accumulate in host cells and theiraction is sustained for a longer time.

1. (canceled)
 2. The method of claim 16, wherein the porous silicateand/or the aluminosilicate matrix is at least one selected from thegroup consisting of MCM-41 and KIT-6.
 3. The method of claim 2, whereinthe nanocarrier has a surface area of 225 to 750 m²/g, a pore volume of0.25 to 0.85 cm³/g, and a mean pore size of 2 to 10 nm.
 4. The method ofclaim 2, wherein the porous silicate and/or aluminosilicate matrix isMCM-41 and the nanocarrier has a surface area of 450 to 750 m²/g, a porevolume of 0.25 to 0.65 cm³/g, and a mean pore size of 2 to 4 nm.
 5. Themethod of claim 2, wherein the porous silicate and/or thealuminosilicate matrix is KIT-6 and the nanocarrier has a surface areaof 225 to 450 m²/g, a pore volume of 0.45 to 0.85 cm³/g, and a mean poresize of 5 to 10 nm.
 6. The method of claim 16, wherein the poroussilicate and/or the aluminosilicate matrix is present in an amount of 1to 20 wt % based on a total weight of the nanocarrier.
 7. The method ofclaim 16, wherein the metal organic framework comprises an imidazolemoiety of formula I:

and is substantially free of a benzimidazole moiety of formula II:


8. The method of claim 7, wherein the zeolitic imidazolate framework isZIF-8.
 9. The method of claim 16, wherein the antioxidant is at leastone selected from the group consisting of quercetin, rutin, coenzymeQ10, gallic acid, resveratrol, and curcumin.
 10. The method of claim 9,wherein the antioxidant is curcumin.
 11. The method of claim 9, whereinthe antioxidant is resveratrol.
 12. The method of claim 16, wherein theantioxidant is present in the nanomedicinal composition in an amount of5 to 50 wt %, based on a total weight of nanomedicinal composition. 13.(canceled)
 14. (canceled)
 15. The method of claim 16, wherein thenanomedicinal composition is capable of reducing the viability ofBlastocystis organisms by at least 75% when the nanomedicinalcomposition is present for 24 hours in an amount of 100 to 1000 μg/mL.16. A method of forming a nanomedicinal composition, the methodcomprising: mixing a metal organic framework and a porous silicateand/or an aluminosilicate matrix to form a nanocarrier; combining thenanocarrier and a antioxidant in an impregnation solution therebyforming the nanomedicinal composition; and isolating the nanomedicinalcomposition; wherein the metal organic framework is a zeoliticimidazolate framework; and the antioxidant is disposed in the poresand/or on a surface of the nanocarrier.
 17. The method of claim 16,wherein the mixing comprising ultrasonication.
 18. The method of claim16, wherein the impregnation solution comprises an alcohol having 1 to 5carbon atoms and the antioxidant is present in an amount of 0.5 to 3mg/mL of impregnation solution. 19-20. (canceled)